IJRPC 2017, 7(4), 407-424 Mounica Ponugoti ISSN: 22312781
INTERNATIONAL JOURNAL OF RESEARCH IN PHARMACY AND CHEMISTRY
Available online at www.ijrpc.com Review Article
A PHARMACOLOGICAL AND TOXICOLOGICAL REVIEW OF MATCHLESS HERB: TULASI Mounica Ponugoti Department of Pharmacology, Hindu College of Pharmacy, Guntur - 522002, Andhra Pradesh, India.
ABSTRACT Nature has many useful herbs and plants for human beings. A majority of world’s population in developing countries still relies on herbal medicines to meet its health needs. Among the plants known for medicinal value, the plants of genus Ocimum belonging to family Lamiaceae are very important for their therapeutic potentials. Ocimum sanctum Linn. (Tulasi), a sacred and traditional medicinal plant of India which possesses innumerable health benefits and therefore regarded as the “Elixir of Life”, ‘Incomparable one’ ‘Machless one’ and ‘Queen of Herbs’. Many research and studies suggest that Tulasi may be a COX-2 inhibitor, like many modern painkillers, due to its significant amount of eugenol. Bioactive compounds of Tulasi responsible for its various medicinal properties and their effects at the molecular level need to be investigated in more detail for pharmaceutical therapeutic applications. The present review summarizes the comprehensive information concerning pharmacological activities such as analgesic, anti-inflammatory, antipyretic, antiulcer, antiarthritic, antiasthamatic, antibacterial, antifungal, antiviral, antihelmintic, antiplasmodic, larvicidal, antioxidant, antistress, anticancer, anticataleptic, anticataract, wound healing, anticoagulant,antioxidant, antidepressant, anticonvulsant, antidiabetic, antifertility, antihyperlipidemic, cardioprotective, antihypertensive, antitussive, antiemetic, genoprotective, hepatoprotective, diuretic, Immunomodulatory, neuroprotective, radioprotective activites along with toxicological studies.
Keywords: Tulasi, Ocimum sanctum, Lamiaceae, Pharmacological activities, Toxicological studies.
INTRODUCTION medicine appeared in Rigveda which has been Medicinal plants are the richest bio-resource of written 3500 - 1600 B.C3. drugs for traditional systems of medicine, Properties of plants as a source of medicine modern medicines, nutraceuticals, food were studied in detailed in Ayurveda, a system supplements, folk medicines, pharamaceutical of traditional Hindu medicine which is native to intermediates and chemical entities for India and is renowned as one of the major synthetic drugs1. Medicinal plants have a very systems of alternative and complementary rich sources of secondary metabolites and oils medicine. According to Hindu mythology, which are of therapeutics importance. Some of Dhanvantari, the physician of the God‟s, is the most important bioactive phytochemical attributed with the origin of ayurvedic constituents in plants are alkaloids, flavonoids, medicine. Ayurveda traces its origin to the phenolics, essential oils, tannins and Vedas particularly Atharvaveda and it stresses saponins. The important advantages of the use of indigenous plant based medicines medicinal plants in various treatments are: for the treatment of diseases4. their safety besides being less expensive, Tulsi “Queen of herbs” is described as sacred efficacy and availability through out the world2. and medicinal plant in ancient literature. It is Use of plants as a source of medicinal value is an important symbol of the Hindu religious a very old concept. In India use of plants as a tradition. The name Tulsi is derived from „Sanskrit‟, which means “matchless one”5. Its
407 IJRPC 2017, 7(4), 407-424 Mounica Ponugoti ISSN: 22312781 other name, Vishnupriya means the one that similar. Both the varieties also have common pleases Lord Vishnu. This plant belongs to the medicinal properties7. family Labiatae, characterized by square stem There are many species of Ocimum, which and specific aroma. Botanical name of Tulsi is have their different morphological or Ocimum sanctum (Linn). In India, the plant is anatomical characters. They are found in grown throughout the country from Andaman different places and have different living and Nicobar islands to the Himalayas up to conditions; so that they have different 1800 meters above the sea level6. It is also medicinal value. Content of abundantly found in Malaysia, Australia, West secondarymetabolites also differs species to Africa and some of the Arab countries. species. Different species of Ocimum are Ocimum sanctum (Linn) is the most prominent Ocimum americanum, Ocimum species of the genera. The leaves of the plant basilicum,Ocimum campechianum, Ocimum are considered to be very holy and often form centraliafricanum, Ocimumgratissimum, a consistent part of the Hindu spiritual rituals Ocimum kilimandscharicum, Ocimum (Tirtha or Prasada). Ocimum sanctum has two minimum,Ocimum viride, Ocimum suave, varieties i.e. black (Krishna Tulsi) and green Ocimum ovatum, Ocimumselloi, Ocimum (Rama Tulsi), their chemical constituents are tenuiflorum and Ocimum citriodorum (O.americanum × O. basilicum)8.
Fig. 1: Plant of Ocimum sanctum (Tulsi)
TAXONOMY Kingdom : Plantae Subkingdom : Tracheobionta Superdivision : Spermatophyta Division : Magnoliophyta Class : Magnoliopsida Subclass : Asteridae Order : Lamiales Family : Lamiaceae Genus : Ocimum Species : O. sanctum
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Morphology color, present in small compact clusters on It is an erect, much branched, fragrant and cylindrical spikes. Stalk less heart-shaped erected plant attaining a height of about 30-60 bracts are there at the base of each flower cm when mature. Its aromatic leaves are cluster. Sepal cup is not hairy within. Flowers simple, opposite, elliptic, oblong, obtuse or are rarely longer than 5 mm, calyx tube acute with entire or sub serrate or dentate bearded outside near base. Flower tube is margins, growing up to 5 cm long. The Tulsi hairy. The fruits are small and the seeds flowers are small having purple to reddish yellow to reddish in colour9.
Table 1: Chemical constituents of O. sanctum Plant Parts Extracts Chemical constituents Aesculectin, Aesculin, Apgenin, Caffiec acid, Chlorgenic acid, Apigenin, Apigenin-o-glucuronide, Triacontanol ferulate, Vicenin-2, Circineol, Gallic acid, Leaves / Alcoholic extract Galuteolin, Isorientin, Isovitexin, Isovitexin, Circineol, Luteolin, Molludistin, areal parts10-13 Orientin, Procatechuic acid, Stigmasterol, Urosolic acid, Vallinin, Viceni, Vitexin, Vllinin acid Vitamin and mineral Vitamin C, Vitamin A, Vitamin E, Calcium, Phosphours, Chromium, Copper, Whole plant 14,11,12 contents Carotene, Zink, Iron, Nickel Aromadendrene oxide, Benzaldehyde, Borneol, Bornyl acetate, Camphor, Caryophyllene oxide, cis-α-Terpineol, Veridifloro, Cubenol, Cardinene, D- Limonene, Eicosane, Eucalyptol, Eugenol, Methyl Eugenol, Farnesene, Leaves 11-17 Farnesol, Furaldehyde, Germacrene, Heptanol, Humulene, Limonene, n- Essential oil butylbenzoate, Ocimene, Oleic acid, Sabinene, Selinene, α-Camphene, α- Myrcene, α-Pinene, β-Pinene, α-Thujene, β-Guaiene, β-Gurjunene, Methyl Chavicol, Linalool, Cirsilineol, Circimaritin phytol, Isothymusin, Apigenin, Rosameric acid, Octane, Nonane, Benzene, Iedol, Cadinene, Borneol
11,12,18,19 Linoleic acid, Linolenic acid, Oleic acid, Palmitric acid, Stearic acid, Sitosterol, Seeds Fixed oil Dilinoleno-linolins, Linodilinolin, Hexoureic acid Whole plant 20 Secondary Alkanoids, Steroids, Tannins, Phenol compounds, Flavonoids, Resins, Fatty metabolites acids, Gums
Pharmacological activity inhibition of 500 mg/kg of the tulsi paste was Analgesic Activity found to be 88.15% as that of the response Fresh leaves of tulasi were investigated for observed with 100 mg/kg of indomethacin and analgesic activity using rat tail method. Tulsi showed considerable anti-inflammatory showed an increase of 20.34 per cent with activity23. mild dose, 43.80 percent with moderate dose Anti-inflammatory activity of essential oil and of 51.47 percent with maximum dose at extract of Ocimum sanctum L. leaf (Eugenol) 90 min. after injection. The regression line was studied in wistar rats by using indicated that the analgesic effect remain upto carrageenan induced hind paw edema 3 hours irrespective of dose concentration. method. The extract was administred 100 Analysis of variance revealed that the mg/kg body weight per i.p and the standard analgesic activity of Tulsi was statistically paracetamol was also administered 5 mg/kg significant with all the three doses [p body weight per i.p. The extracted Eugenol 21. and paracetamol exhibited significant (p< The analgesic activity of fixed oil from the 0.05%) activity when compare with seeds of Ocimum sanctum (OS) were carrageenan control24. investigated in mice and rats using the tail Different extracts of stem, leaf and stem calli flick, tail clip, tail immersion and acetic acid- of OS were tested for antiinflammatory activity induced writhing methods. It was found it be using carrageenaninduced rat paw oedema effective against acetic acid induced writhing model in comparison with the standard in dose dependent manner, suggesting that indomethacin. The ethanol extract of callus writhing inhibiting activity of the oil is tissue exhibited maximum significant anti- peripherally mediated due to combined inflammatory activity out of all extracts studied inhibitory effects of prostaglandins, histamine followed by ethanol extracts of leaves of OS25. and acetylcholine22. Ocimum sanctum fixed oil and linolenic acid were found to possess significant Antiinflammatory Activity antiinflammatory activity against PGE2, The fresh tulsi leaf in its paste form was leukotriene and arachidonic acid-induced paw tested for anti-inflammatory activity using edema. Plant lipids like linseed oil and carrageenan induced paw edema model in soyabean oil containing linolenic acid when comparison to Indomethacin. The percent tested along with OS fixed oil, also showed
409 IJRPC 2017, 7(4), 407-424 Mounica Ponugoti ISSN: 22312781 significant inhibition of carrageenan-induced significant protection against ethanol induced paw edema. The results suggest that linolenic gastric ulceration in Wistar rats. OS exhibits acid present in OS O fixed oil has the capacity antiulcer activity by enhancing antioxidant to block both the cyclooxygenase and potential of gastric mucosa there by reducing lipoxygenase pathways of arachidonate mucosal damage31. metabolism and could be responsible for the It was found that the ethanolic extract of OS antiinflammatory activity of the oil26. not only reduced acid secretion, but also Experimental studies of Tulsi have shown to potentially elevated the mucoprotective effect inhibit acute as well as chronic inflammation in and 100 mg/kg body weight was found to be rats. This test was conducted by carrageenan the most effective dose in dose dependent induced paw edema, croton oil induced manner indicating that OS extract exhibited granuloma, and exudates at a dose of 500 antiulcerogenic in all the five models agaisnt mg/kg, bw/day. The oils processed from fresh ulcer induced by cold restraint (CRU), alcohol leaves and seeds of O. sanctum have (AL), aspirin (ASP), and pyloric ligation (PL) revealed anti-inflammatory effects on model in rats, and histamine (HST) induced experimental animals induced by carrageenan, duodenal ulcer model in guinea pigs32. histamine, serotonin and prostaglandin E2 The fixed oil of OS administered i.p. shows according to some studies. These significant antiulcer activity against aspirin, experimental rats were administered with indomethacin, alcohol (ethanol 50%), essential oil (200 mg/kg, bw) and fixed oil (0.1 histamine, reserpine, serotonin or stress- ml/kg, bw) before injection of phlogistic agents induced ulcers in rats. The fixed oil and were compared with standard drug significantly possessed antiulcer activity due to flurbiprofen. It was noted that Tulsi extracts its lipoxygenase inhibitory, histamine could significantly reduce the edema antagonistic and antisecretory effects33. whencompared with the saline treated control. Ashok Kumar et al., (2011) while working for However, its effect was less than the standard antiulcer activity of poly herbal formulation drug27. Fixed oil of Tulsi can prevent enhanced (PHF) containing Ocimum sanctum, Abutilon vascular permeability and leukocytic activity as Indicum andTriumfetta Rhomboidea in evidenced by carrageenan induced indomethacin and Ethanol induced ulcers inflammatory stimulus28. showed that PHF has potential antiulcer activity as comparable with standard drugs like Antipyretic Activity Misoprostol (0.012mg/kg) and Omeprazole The antipyretic activity of fresh leaves of tulasi (10mg/kg), at a dose level of 200mg/kg. were evaluated by inducing fever using 15% of brewer‟s yeast suspension, injected Antiarthritic Activity subcutaneously in rats. 18 hrs after giving The fixed oil of Ocimum sanctum seeds was injection, each rat was fed orally with vehicle screened for antiarthritic activity by Singh et al. and test drug accordingly. Temperature was in 1996 using Freund's adjuvant arthritis, recorded to all animals at every 30, 60,120 formaldehyde-induced arthritis and turpentine and 180 minutes respectively. There was a oilinduced joint edema in rats. The fixed oil significant reduction of fever in Tulsi group showed significant anti-arthritic activity in both whereas the temperature control was not models and anti-edema activity against significant statistically in control group. turpentine oil-induced joint edema34. Therefore the crude natural preparation of Tulsi itself, without any processing has Antiasthmatic Activity effective antipyretic action29. 50% aqueous ethanol extract of dried and The antipyretic activity of OS fixed oil was fresh leaves, and the volatile and fixed oils of evaluated by testing it against typhoid OS was evaluated against histamine and paratyphoid A/B vaccine-induced pyrexia in acetylcholine induced preconvulsive dyspnea rats. The oil on ip administration considerably (PCD) in guinea pigs. The 50% ethanol extract reduced the febrile response indicating its and volatile oil extracted from fresh leaves and antipyretic activity. At a dose of 3 ml/kg, the fixed oil from the seeds significantly protected antipyretic activity of the oil was comparable to the guinea pigs against histamine and aspirin. Further, the fixed oil possessed acetylcholine induced pre convulsive dyspnea. prostaglandin inhibitory activity and the same However, the 50% ethanol extract of dried could explain its antipyretic activity30. leaves did not protect the guinea pigs against histamine induced preconvulsive dyspnea27. Antiulcer Activity The aqueous extract of Ocimum sanctum (100mg /kg an 200 mg/kg orally) exhibited
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Antibacterial Activity observed equally effective against pathogenic Biochemical compounds present in methanolic gram positive and gram negative bacteria38. Tulsi leaf extract showing antimicrobial activity Tulsi is known to possess antimicrobial activity against human and fish pathogens were against various bacteria, the most common carried out using Bacillus sp, E.coli, being Candida albicans, Staphylococcus Streptococcus, Bacillus subtilis, Bacillus aureus, Escherichia coli by its cereus, Staphylococcus aureus, Vibrio phytoconstituents isolated from various parts. cholera, Salmonella typhi, Klebsiella In view of this, various studies have been pneumonia, Salmonella paratyphi and Fish conducted, wherein according to Singh et al., pathogens Aeromonas hydrophila, higher content of linoleic acid in O. sanctum L. Pseudomonas aeruginosa, Edwardsiella tarda. fixed oil could contribute toward its Maximum antibacterial activity exhibited antibacterial activity. The oil contains against Bacillus sp. and moderate activity in antibacterial activity against S. aureus, Bacillus subtilis, Bacillus cereus, Vibrio Bacillus pumius, and Pseudomonas cholera, Salmonella typhi. Minimum activity aeruginosa, where S. aureus was the most was noted in Salmonella paratyphi. no sensitive organism39. Similarly Geeta et al. inhibition zone was showed in E.coli., reported that on comparing alcoholic and Streptococcus sp, Staphylococcus aureus, aqueous extract, the aqueous extract of O. Klebsiella pneumonia. Whereas, the fish sanctum L. (60 mg/kg) showed wide zones of pathogens with methanolic crude extract of inhibition against Klebsiella40. tulsi showed significant activity against Antimicrobial activity against pathogens like Aeromonas hydrophila, Pseudomonas Escherichia coli, Staphylococcus aureus, aeruginosa and Edwardsiella tarda35. Bacillus anthracis, Bacillus subtilis, Salmonella Antimicrobial activity of different extracts spp., P. vulgaris, Pseudomonas aeruginosa (Ethanol, Methanol, Ethyl acetate and and Mycobacterium tuberculosis were stuided chloroform) of dried leaf of O. sanctum were and found its activity against E. coli, Klebsiella tested against three human pathogens strains aerogens, Proteus mirabilis, Salmonella such as Escherichia coli, Staphylococcus typhimurium, Shigella dysentriae, Vibrio spp., aureus and Candida albicans through the well P. aeruginosa, cholera and S. aureus41. diffusion and the poison plate method. The Antimicrobial activity was also found against Minimum inhibitory concentration (MIC) values Pasturella multocida, E. coli, S. aureus, B. of the crude extract of the tested plant leaves subtilis and Salmonella typhi, Salmonella were determined. Both methods (well diffusion paratyphi A and Salmonella typhimuriuum and and poison plate) showed the strongest E. coli, Klebsiella spp., B. subtilis, S. aureus 42. activity in methanol extract. Among four In vitro studies against specific pathogens methanol extracts, they show more inhibition suggest that essential oil of Tulsi leaves have against in S. aureus than E. coli and C. significant inhibitory effect against E. coli, B. albican.36. subtilis, B. anthracis, S. aureus, Pseudomonas The antimicrobial activity of Ocimum sanctum vulgaris and P. aeruginosa. These essential leaf extract in normal tap water and local river oils include major constituents of leaves such water was investigated. The antimicrobial as caryophyllene; eugenol, methyl eugenol effect was studied with different concentration which are effective against (100 to 600 mg l-1) of Tulsi leaf extract in tap Arthobacterglobiformis, B. megatherium, E. and river water. In this, 600 mg l-1 coli and Pseudomonas sp43. concentration of plant extract treated water Grover and Rao in 1977 stated that Eugenol is showed effective antimicrobial activity at 15 to the most therapeutically effective constituent 16 hrs than the other concentration of extract. of Tulsi 44. Aqueous and alcoholic extracts of The 500 mg l-1 of extract treated water leaves impart a potentially effective showed 95 to 98% antibacterial activity in 14 antibacterial activity. The extract is effective to 16 hrs. The minimum bacterial against various enteric pathogens viz., E. coli, concentration (MBC) was observed in 500 and K. aerogens, P. mirabilis, Salmonella 600 mg l-1 extract concentration. The typhimurium, Shigella dysentriae, P. concentration of the bacterial cells inhibited aeruginosa, Vibrio cholera and S. aureus. gradually for an hour was studied by spread Antibacterial activities of seeds were also plate method37. studied and it was revealed that the crude, Antibacterial activity of the aqueous, alcoholic, supernatant, residue and dialyzed samples chloroform extract and oil obtained from obtained from the seeds inhibited the growth leaves of Ocimum sanctum were studied of P. multocida, E. coli, B. subtilis and S. against E.coli, P.aeruginosa, S. typhimurium aureus45,46. and S.aureus. Extract obtained from OS were
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Growth inhibition of Klesbiella, E. coli, and lesions of the plasma membrane and a Proteus and S. aureus by aqueous extract was considerable reduction in the amount of studied. On the other hand, the aqueous ergosterol53. extract has activity against the notorious Antifungal activity of Ocimum sanctum leaves multidrug-resistant strains of S. aureus which was determined against clinically isolated show resistance to beta lactam antibiotics. O. dermatophytes. Minimum inhibitory sanctum is also active against resistant strains concentration (MIC) and minimum fungicidal of Neisseria gonorrhea, the fixed oil has an concentration (MFC) of various extracts and efficient good antibacterial activity against fractions of OS leaves were also determined Bacillus pumilus, P. aeruginosa and S. aureus. against dermatophytic fungi used54. Linoleic acid also has antibacterial activity. In addition to antibacterial the essential oil also Antiviral Activity has insecticidal properties. It has ten times the Different types of extracts of Ocimum sanctum anti-tubercular potency of streptomycin and have anti-viral activity against different viruses approximately one-fourth times the activity that e.g. Hematopoietic Necrosis Virus (IHNV)55, of isoniazid. The essential oil is effective polio virus type 356, herpes virus (HSV), against pathogenic fungi including Alternaria hepatitis B virus, New castle Disease Virus. solani, Candida guillermondii, Colletotricum Ethanolic extract of Tulsi plant leaves in a capsici, Curvularia spp., Fusarium solani, range of 22.5 mg/ml concentration inhibit Helminthosporium oryzae and the bacterial replication of polio type 3 virus in VERO cells. strains, Anthrobacter globiformis, Bacillus The extracted components of this plant like megaterium, E. coli, Pseudomonas spp., S. linalool, apigenin and ursolic acid show broad aureus, S. albus and Vibrio cholerae 47. spectrum antiviral activity against DNA viruses The essential oil has activity against both like RNA virus and adenoviruses57,58. One Gram-positive as well as Gram-negative study also proves its efficacy against new bacteria. For enteric pathogens, aqueous castle disease of poultry 59. extract and alcoholic extract is beneficial while Tulasi is used in most of the countries on the contrary, seed oil of Tulsi yields worldwide to help protect against swine flu. considerable antimicrobial properties. The The main chemical constituents isolated from ethanolic extract inhibits methicillin-resistant S. leaves are Ursolic acid, apigenin and luteolin. aureus (MRSA) which is notorious for the Several formulations are available in the production of B-lactamases and significant market60,61. Boiled tulsi that is basil leaves activity is also demonstrated against served in a warm drink like tea can help methicillin-sensitive S. aureus [MSSA]. The prevent an outbreak of dengue. This bitter and oils have antimicrobial activity against Propioni pungent herb has all the properties that bacterium acnes. It has minimum inhibitory strengthen the internal system against fever62. concentration (MIC) of 3.0% v/v. Viral encephalitis patients benefit from aqueous Anti-helminthic Activity extract. Tulsi leaves paste was found effective The essential oil of Ocimum sanctum and against ring worm infections. Tulsi naturally eugenol, tested in vitro, showed potent possesses antimicrobial properties and is used anthelmintic activity in the Caenorhabditis in the treatment of many serious systemic elegans model63. diseases and localized infection. With fresh juice and honey, worms and parasites are Antiplasmodial Activity removed; the sweetness excites the parasites Leaf extract, root extracts, the stem and flower out. It is used in the treatment of viral extracts of OS showed excellent encephalitis, malaria and typhoid48. antiplasmodial activity in a study carried out by Tulsi demonstrated effective antimicrobial Inbaneson et all in 2012 on three different property against Aggregatibacter species of ocimum. The in vitro antiplasmodial actinomycetemcomitans, suggesting its activity might be due to the presence of possible use as an effective and affordable alkaloids, glycosides, flavonoids, phenols, “adjunct” along with the standard care in the saponins, triterpenoids, proteins, resins, management of periodontal conditions49-52. steroids and tannins in the ethanolic extracts of tested plants64. Antifungal Activity Methyl chavicol and linalool obtained from Larvicidal Activity essential oil of Ocimum sanctum showed Larvicidal activity of essential oils and different significant antifungal activity against Candida, extracts of Ocimum. Sanctum, O. basilicum including azole-resistant strains. Their and O. gratissimum were compared on fungicidal action resulted from extensive laboratory reared and field collected larvae of
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Culex quinquefasciatus. The LD50 value of O. leaves of O. sanctum. The anticancer activity basilicum and O. sanctum oil were 39.31 and of Tulsi has been reported against human 40.02 on laboratory reared larvae and 129.53 fibrosarcoma cells culture, wherein AlE of the and 139.49 on field collected larvae. drug induced cytotoxicity at 50 mg/ml and Laboratory reared larvae were more sensitive above. In such studies, microscopically, the than field collected larvae.45 The acetone, cells showed shrunken cytoplasm and chloroform, ethyl acetate, hexane, and condensed nuclei. The DNA was found to be methanol leaf and flower extracts of Ocimum fragmented when observed in agarose gel sanctum were studied against fourth instar electrophoresis70. larvae of Aedes aegypti and Culex Antimelanoma activity of 50% alcoholic quinquefasciatus. The highest larval mortality aqueous leaf extract of different species of was found in leaf extract of O. sanctum Ocimum were investigated. Leaf extract against the larvae of aegypti and C. administered orally (200mg/kg, p.o.) resulted quinquefasciatus65. in significant reduction in tumor volume, increase in average body weight, and survival Antioxidant Activity rate of mice71. The antioxidant effects of Ocimum sanctum The seed oil of Ocimum sanctum was were investigated in experimental evaluated for anticancer activity against streptozocin-induced diabetic rats. subcutaneously injected 20- Administration of OS to streptozocin-induced methylcholanthrene induced-fibrosarcoma diabetic rats for 30 days significantly reduced tumors in the thigh region of Swiss albino the plasma level of thiobarbituric acid reacting mice. The enhanced survival rate and delay in substances and improved the status of the tumor incidence was observed in seed oil antioxidant enzymes catalase, superoxide supplemented mice. Potential dismutase and glutathione peroxidase in vital chemopreventive activity of the oil is partly organs such as the liver and kidney66. attributable to its antioxidant properties. The A hydroalcoholic extract of OS leaves has chemopreventive efficacy of 100 microl/kg been investigated for its antioxidant activity in seed oil was comparable to that of 80 mg/kg of animal models of peptic ulcer with the aim of vitamin E72. exploring a possible correlation between its Papilloma genesis induced by 7,12- antioxidant and antiulcer activities. The dimethylbenz(a) anthracene (DMBA) antioxidant activity was by evaluated by significantly reduced the tumor occurrence in estimating plasma malondialdehyde (MDA) in mice on topical application of O. sanctum leaf ethanol treated rats and histamine treated extract. The application of Tulsi extracts in the guinea pigs and estimating superoxide form of paste has shown promising results in dismutase (SOD) in pyloric ligated rats and the prevention of DMBA induced buccal pouch histamine treated guinea pigs. In ethanol carcinogens73. treated rats and histamine treated guinea pig Different types of carcinogens have been tried ocimum sanctum leaf extract (100 mg/kg & for evaluating the anticarcinogenic properties 200 mg/kg) significantly decreased the levels in the experimental animals induced by Tulsi of MDA in comparison the diseased control. leaves when fed to experimental rats with 600 The extract (100 mg/kg & 200 mg/ kg) also mg/g diet for 10 weeks, significantly reduced increased the levels of SOD in pyloric ligated the 3,4-benzo (a) pyrene [B (a) P] and 3„- rats and histamine treated guinea pigs when methyl-4- dimethylaminoazobenzene compared to the diseased control67. (3„MeDAB)- induced squamous cell carcinoma and hematoma incidences74. Antistress Activity Administration of 70% ethanolic Tulsi leaf Fresh leaves of Ocimum sanctum were extract has also been observed to reduce the evaluated for antistress activity against incidence of cancer caused by N-methyl-N′- experimentally induced oxidative stress in nitro-N-nitrosoguanidine (MNNG), a nitroso albino rabbits68. compound widely used as an experimental gastric carcinogen. MNNG is a potent Anticancer Activity mutagen and induces erosions of the gastric Tulsi has been shown to possess an excellent mucosa, an initial precancerous change anticancer activity69. Detoxification of integral for the initiation of stomach carcinogens and mutagens which is carried carcinogenesis. Intragastric administration of out by enzymes such as glutathione-S- MNNG induces increased cell proliferation and transferase, cytochrome b5 and cytochrome angiogenesis with evasion of apoptosis P450, and aryl hydrocarbon hydroxylase is leading to well differentiated squamous cell modulated by the alcoholic extract (AlE) of carcinomas. Administration of Tulsi has been
413 IJRPC 2017, 7(4), 407-424 Mounica Ponugoti ISSN: 22312781 shown to decrease these activities wherein Anticataract Activity Tulsi extract influences the critical molecules The Aqueous Extract of fresh leaves of OS involved in cell proliferation, invasion, (1g/kg and 2 g/kg) significantly delayed the the angiogenesis, and apoptosis. A significant onset as well as subsequent maturation of decrease in the levels of cytokeratin, CK cataract in galactosemic cataract model in rats (infiltration), vascular endothelial growth factor, by 30% galactose and naphthalene cataract VEGF (angiogenesis), proliferating cell nuclear model in rabbits by 1 g/kg naphthalene80. antigen (PCNA), glutathione-S-transferase pi (key proteins involved in proliferation), and Wound healing antiapoptotic protein Bcl-2 with simultaneous Wound healing activity of Ocimum sanctum is increase in the proapoptotic proteins Bax, also proved by using two different types of cytochrome c, and caspase 3 were reported75. concentration (200 and 400 mg/kg) in rats. Studies also suggest that the leaf extract The models of wound used for this study are: blocks or suppresses the biochemical events the excise, the incise and dead space wound associated with chemical carcinogenesis by model. By using Van Gieson and Masson preventing metabolic activation of the Trichome strains in histological examination of procarcinogen to carcinogen. Previous studies determination of granuloma tissue, it is found suggested that AlE of Tulsi leaf before that Ascorbic acid, Hexose amine, L- administering 7,12 dimethylbenz[a] anthracene Hydroxyproline and Malondialdehyde isolated causes decreased phase I enzymes; reduction from Tulsi has wound healing activity. Tulsi in the levels of lipid and protein oxidation, and can be used as adjunct therapies for the burn a concomitant enhancement of the antioxidant wound management many studies supporting and phase II enzyme activities in the liver. its use in healing81-83. Tulsi also causes a decrease in the 7,12- dimethylbenz[a] anthracene induced Anticoagulant Activity genotoxicity, as evaluated by the micronuclei Ocimum sanctum fixed oil (3 ml/kg, ip) was formation in bone marrow cells in mice. These studied for anticoagulant activity. It was results suggest that, in association with the observed that blood clotting time was modulation of the phase I and II detoxification prolonged and the response was comparable enzymes, Tulsi possesses antigenotoxic to that obtained with aspirin (100 mg/kg). The effects, and all these might have contributed to effect appears to be due to the anti- the reduction of chemical carcinogenesis76. aggregatory action of oil on platelets84. Tulsi decreased the expression of cutaneous γ-glutamyl transpeptidase (GGT), a marker of Antianxiety and Antidepressant Activity tumor progression, and glutathione-S- The effect of ethanol extract of leaves of transferase-P, which is increased in chemically Ocimum sanctum in Swiss albino mice, induced hepatic tumors. The heat shock against both anxiety and depressive disorders protein, which is altered during were investigated. Depression was studied carcinogenesis, has also shown a decrease in through tail suspension test and forced swim its concentration77. test. Anxiety experiments included light dark Application of Tulsi extract decreased the test, elevated plus maze test, and holeboard activity of ornithine decarboxylase, an enzyme test. The Ocimum sanctum extracts shows involved in the regulation of cell proliferation antianxiety and antidepressant properties at and development of cancer. There was also a the same dose and can be a potential concomitant decrease in the phase I enzymes therapeutic agent against mixed anxiety and and lipid peroxidation suggesting that O. depressive syndrome85. sanctum prevents the activity of carcinogen induced cytochrome P-450 dependent Anticonvulsant Activity enzymes and that this leads to a decrease in Different extractives of stem, leaf and stem the formation of ultimate carcinogenic callus of Ocimum sanctum were tested for moiety78. anticonvulsant activity against standard drug phenytoin using maximal electroshock (MES) Anticataleptic Activity model. Ethanol and chloroform extractives of The anticataleptic activity of the aqueous stem, leaf and stem calli were effective in extract (300 mg/kg, i.p) and the alcoholic preventing tonic convulsions induced by extract (300 mg/kg, i.p) of the leaves of transcorneal electroshock86. Ocimum sanctum was studied and observed a significant (P < 0.001) reduction in cataleptic Antidiabetic Activity scores79. Ten fractions (F1-F10) were isolated from hydroalcoholic extract of OS aerial part by
414 IJRPC 2017, 7(4), 407-424 Mounica Ponugoti ISSN: 22312781 column chromatography. All the fractions F1 to marked increase while FSH and LH levels F10 were screened for antidiabetic activity in were significantly reduced in OS-treated alloxan induced diabetic rats by estimating rabbits (2 g fresh leaves/rabbit for 30 days). serum glucose level and lipid parameters. The The results suggest the potential use of OS as bioactive fraction (F5) was found to be potent an effective male contraceptive agent91. antidiabetic by ameliorating glucose and lipid parameters (total cholesterol, triglycerides, low Antihyperlipidemic and Cardioprotective and high density lipoprotein cholesterol). The Activity extensive spectroscopic data analysis reveals The antihyperlipidemic and cardioprotective that, the isolated bioactive compound activity of Ocimum sanctum fixed oil was elucidated as tetracyclic triterpenoid87. studied in rats fed with a high fat (HF) diet and The effect of ethanolic extract and five partition concluded that treatment with OS fixed oil fractions of OS leaves on insulin secretion decreased the high serum lipid profile and together with an evaluation of their expressed antiartherogenic and mechanisms of action were studied and cardioprotective actions against concluded that Ocimum sanctum leaf extracts hyperlipidemia. The anti-hyperlipidemic action stimulate insulin secretion from perfused of OS fixed oil was mainly resulted from the pancreas, isolated islets and clonal suppression of liver lipid synthesis. Linolenic pancreatic–cells88. acid and linoleic acid contained in Ocimum The antidiabetic effects of Ethyl acetate, sanctum fixed oil were possibly responsible for Petroleum-ether, and Chloroform fractions both lipid-lowering and cardiac protective from ethanolic extract of the leaves of OS action against hyperlipidemia92. were investigated in normal and alloxan In Streptozotocin induced diabetic rats, induced diabetic rats (AIDRs). Administration aqueous extract of tulsi was administered for of these fractions to the AIDRs resulted in the eight weeks and decrease in lipid profile was significant elevation of liver glycogen content. observed93. In diabetic rats, SGOT and SGPT levels were significantly elevated that were further reduced Antihypertensive Activity after i.p. administration of these fractions. The OS fixed oil administered i.v. produced These results indicate that different fractions of hypotensive effect in anaesthetized dog, which OS have favorable effects in bringing down the seems to be due to its peripheral vasodilatory severity of diabetes together with action. Essential fatty acids like linoleic and hepatoprotectivity89. linolenic acids, contained in the OS oil produce Methanolic extracts of leaves of various series 1 and 3 (PGE1 and PGE3) Ocimum species were explored and compared prostaglandins and inhibit the formation of for antidiabetic activity. All extracts were able series 2 prostglandins (PGE2)84. to show antidiabetic activity at 0.5 mg/Kg concentration. The activities are well Antitussive Activity comparable with the standard drug, Aqueous and methonolic extract of Ocimum glibenclamide. The methanolic extract of OS sanctum was studied for antitussive activity in showed better antidiabetic activity in guinea pigs at the doses of 1.55 gms and comparison with other species of Ocimum and 0.875 gms/kg body wt respectively. Cough standard drug. The data were verified as was induced by exposure to the aerosol of statistically significant by using one way citric acid (7.5% w/v). The study showed that ANOVA at 5 % level of significance (p < both the test extracts posses significant 0.05)90. antitussive activity and aqueous extract showed a higher activity than the methonolic Antifertility activity extract94. Treatment of albino rats with a benzene extract of Ocimum sanctum leaves (250 mg/kg Antiemetic Activity body weight) for 48 days decreased total Tulsi leaves also check vomiting and used for sperm count, sperm motility, and forward antiemetic action95. velocity. The results suggest that such effects are due to androgen deprivation, caused by Antithyroidic Activity the anti-androgenic property of OS leaves. Effects of Ocimum sanctum leaf extract was The effect was reversible because all investigated on the changes in concentrations parameters returned to normal 2 week after of serum T3, T4 in the male mouse. OS leaf the withdrawal of treatment.24 A significant showed anti-thyroidic activity96. decrease was noted in the sperm count in rabbits. Serum testosterone levels showed
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Eye Disease statistically significant. Ocimum sanctum The leaf juice of Ocimum sanctum along with extract significantly increased the volume of triphla is used in ayurvedic eye drop urine (5.48±0.13ml/100g/24hr and preparations recommended for glaucoma, 7.52±0.19ml/100gm/24hr), increasing the chronic conjunctivitis & other painful eye diuretic index to 1.65 and 2.26 for 250mgkg disease. In daily routine one may use about and 500mg/kg dose ranges respectively (P< three drops of tulsi oil alog with honey and it is 0.01). The test drug, when compared to the supposed to improve eye sight97. control group, showed a significant increase in the excretion of sodium, potassium and Genoprotective Activity chloride excretion. There was an increase in Protective effect of Ocimum sanctum was the saluretic index as reflected by the Na/K evaluated on chlorpyrifos-induced genotoxicity ratio to 2.2 and 2 respectively for the two in in vivo and in vitro models. It was observed dosages studied when compared to frusemide that rats pretreated with OS extract, showed a which showed a saluretic index of 1.81. These significant (P<0.01) increase in mitotic index a findings support the use of Ocimum sanctum significant decrease in the frequency of as a diuretic agent with an action similar to aberrant cells as compared to the rats treated that of the loop diuretic, frusemide101. with chlorpyrifos alone. A significant (P<0.05) increase in chromosomal aberrations was Immunomodulatory Activity observed in cultures treated with 75 μg/ml The aqueous extract of Ocimum sanctum at chlorpyrifos as compared to controls, which the oral doses of 100, 200 mg/kg/day in rats decreased significantly (P<0.05) with Ocimum enhanced the production of RBC, WBC, sanctum extract pretreatment98. haemoglobin and also enhanced the production of antibodies without affecting the Hepatoprotective Activity biochemical parameters102. The hepatoprotective activity of Ocimum Modifications in the humoral immune response sanctum alcoholic leaf extract was studied in rats was observed when treated with against paracetamol-induced liver damage in distilled extract of fresh leaves attributing to Albino rats synergism with silymarin and mechanisms like antibody production, tissue concluded that Ocimum sanctum alcoholic leaf responses, release of mediators of extract showed significant hepatoprotective hypersensitivity in specific organs. Seed oil activity and synergism with silymarin99. was observed to regulate both cell mediated When alcoholic extract of Tulsi plant orally and humoral immune response. The GABA administered, it exhibited hepatoprotective pathways may demonstrate the effect against Paracetamol, Carbon immunomodulatory effects. Tulsi enhances tetrachloride and anti-tuberculosis drugs both cellular and humoral immunity103. induced liver injury in albino rats. When extract The aqueous extract of leaf had of Ocimum sanctum were used in male albino immunotherapeutic potential in sub-clinical rats weighing 100-150 g of Wistar strain (5-6 trials of bovine during intra-mammary aqueous weeks) the level of enzymes was reduced. extract infusion and it was also stated that Biometry Research Unit, Indian Statistical Ocimum sanctum L. aqueous extract produces Institute, 203 revealed that cold water extract a reduction in the bacterial total count and an of Tulsi plant produced hepatotonic effect increase in the count of neutrophil and against Paracetamol and Carbon tetrachloride lymphocyte and demonstrated a good when albino rats fed orally for 6 days with Tulsi phagocytic ability104. extract100. The immunomodulatory effects produced by O. sanctum L. seed oil was studied in both Diuretic activity non-stressed as well as stressed animals for The diuretic activity of aqueous extract of some immunological parameters. Ocimum sanctum was investigated in healthy Consequently, it was stated that Tulsi seed Wistar albino rats. The study was conducted in regulates both humoral and cell-mediated saline primed Wistar albino rats (n=6) using immune responses mediated by GABAergic frusemide (20 mg/kg per oral) as the reference pathway. Godhwaniet al. checked the diuretic drug with two oral doses of ethanolic immunoregulatory effect demonstrated by both extract of Ocimum sanctum (L.) 250mg/kg and methanolic extract along with an aqueous 500mg/kg respectively. Urine volume and suspension of Tulsi leaves for the treatment of electrolytes (Sodium, Potassium and Chloride) antigenic challenge provoked by Salmonella excretion was estimated at the end of 24 typhosa together with sheep erythrocytes and hours. Data was analyzed by ANOVA followed to quantify antibodies that had been by Tukey‟s test. P < 0.05 was considered as agglutinating by Widal agglutination and sheep
416 IJRPC 2017, 7(4), 407-424 Mounica Ponugoti ISSN: 22312781 erythrocyte agglutination tests and in albino rats. Maximal electroshock, atropine, and rats. The results indicated an immune cyclosporine were used to induce dementia. stimulation of humoral immunogenic response The passive avoidance taskwas used for due to increased antibody titer in the Widal assessing memory. Acetylcholinesterase together with sheep erythrocyte agglutination (AChE) activity was estimated in different parts tests103,105,106. of the brain, and immune status was studied Tulsi is an effective immunomodulatory plant. using dinitrochlorobenzene (DNCB) skin Modification in the humoral immune response sensitivity tests. In all the three models both was observed by distilled extract of fresh aqueous and alcoholic OS extracts decreased leaves. Aqueous extract of leaves in vitro the time taken to reach the shockfree zone showed that leaves had proliferative as well as and the number of mistakes and significantly inhibitory effect on splenocytes. In comparison decreased the AChE activity in rats. OS to negative control, 42.17, 55.42 and 47.38% treatment significantly increased the induration increasein the proliferation of spleen cells in the DNCB skin test. Therefore, OS was were reported when splenocytes culture was shown to be useful for the management of treated with 31.25, 62.5 and 125 μg/ml Hot experimentally induced cognitive dysfunctions aqueous extract of O. sanctum In comparison in rats110. to positive control, spleen cells with Hot The alcoholic extract of dried whole plant of aqueous extract of O. sanctum leaves in OS ameliorated the amnesic effect of presence of Con-A exhibited 1.25 and 12.36% scopolamine (0.4 mg/kg) and aging-induced increase in the proliferation of spleen cells memory deficits in mice. Passive avoidance when splenocytes culture was treated with paradigm served as the exteroceptive 31.25 μg/ml and 62.5 μg/ml HAE of O. behavioural model. OS extract increased step- sanctum, respectively. The methanolic extract down latency (SDL) and acetylcholinesterase together with an aqueous suspension of O. inhibition significantly111. sanctum leaves produced clinically evident Various behavioural tests and biochemical immunostimulation of humoral immunological were performed to explore the possible role of response107. OS in alzheimer‟s disease.OS exhibited A combination of O. sanctum, ascorbic acid anxiolytic activity in open field test. In elevated and verapamil were given to experimental plus maze test OS significantly allevated animals exposed to cocaine, they enhanced ibotenic acid and colchicine induced anxiety the macrophage function and decrease and depression in Porsolt‟s swim test. In oxidative stress. Aqueous and ethanolic Morris‟ water maze test, OS pretreatment extract of leaves was used to study improves reference memory, working memory immunomodulatory activity on specific and and spatial learning. Both ibotenic acid and nonspecific immunity in mice, that show colchicine induced deficits in active avoidance strengthening of both specific and non-specific learning and retention of learned behavior responses that can be assessed with were significantly reversed. OS might be haemagglutination antibody (HA) titer, effective in clinical Alzheimer‟s disease by neutrophil virtue of its cognition enhancement, adhesion test, Delayed Type Hypersensitivity antidepressant and antianxiety properties, (DTH)106,108. which are primary needs to be addressed in alzheimer‟s disease112. Neuroprotective Activity Ocimum sanctum shows ameliorative potential Radio-protective Activity in attenuating vincristineinduced peripheral Joseph et al., in 2011 studied the neuropathic pain in rats, which may be radioprotective effect of Ocimum sanctum on attributed to decrease in oxidative stress and the salivary gland of rats administered calcium levels. Administration of OS (100 and radioiodine ((131)I) and compared its efficacy 200 mg/kg p.o.) and its saponin rich fraction with a known radioprotectant, amifostine. OS (100 and 200 mg/kg p.o.) for 14 days and amifostine presupplemented and significantly attenuated vincristine-induced subsequently exposed to (131)I rats at 3 and 6 neuropathic pain along with decrease in months duration exhibited comparable oxidative stress and calcium levels109. histopathology with controls. The study indicated possible radioprotective effect of OS Memory Enhancer Activity and amifostine against high-dose (131)I Aqueous (300 and 500 mg/kg) and alcoholic exposure113. (300 and 500 mg/kg) extracts of Ocimum Flavonoids extracted from the leaves of, OS sanctum Linn. Leaves were studied for were studied as a raddioprotector on the antidementic and anticholinesterase effect in erythrocyte antioxidants in oral cancer. Results
417 IJRPC 2017, 7(4), 407-424 Mounica Ponugoti ISSN: 22312781 of the study suggest that erythrocytes from result showed that Ocimum sanctum extract cancer patients responded to oxidative stress could be safe for human use120. by elevating glutathione levels, while a On administration by oral route, approximate decrease in glutathione levels observed in OS LD50 of Ocimum sanctum was found to be flavonoids treated patients , could be due to 4505±80 mg/kg body weight(bw) and by intra- the free radical scavenging effect of OS peritoneal (ip) routes, 3241±71 mg/kg, bw. OS flavonoids, sparing the glutathione. However leaves aqueous and alcoholic extracts with OS flavonoids did not seem to exert its effect graded doses (3500–6300 mg/kg, bw) were on other antioxidants of erythrocytes114. injected ip in mice, and after a period of 72 hours it was observed that aqueous extract Antidote activity administration at doses up to 5 g/kg body OS showed antidote activity to many poisons. weight did not produce any toxic effect ie OS can be used antidote for dog bite, scorpion 100% safe while 80% tolerance was shown by bite, snake bite and insect bites115-117. alcoholic extract up to a dose of 4g/kg, bw. The acute LD50 (30) values for aqueous Toxicity Studies extract was found to be 6200 mg/ kg, bw while The median lethal dose (LD50) of OS fixed oil that of alcoholic extract was found to be 4600 was determined after ip administration in mice. mg/kg, bw121. The fixed oil was well tolerated up to 30 ml/kg, while 100% mortality was recorded with a CONCLUSION dose of 55 ml/kg. The LD50 of oil was 42.5 It is evident that Tulsi is a medicinal plant of ml/kg. There was found no untoward effect on great importance because of its varied subacute toxicity study of OS fixed oil at a application in medicine, and hence can be dose of 3 ml/ kg/day, ip for 14 days in rats118. corroboratively called the “Queen of Herbs.” . Acute toxicity studies of leaves powder of Several medicinal properties have been ocimum plant material was carried out in swiss attributed to the plant not only in Ayurveda and mice weighing 25-35 gms by administering a Siddha but also in Greek, Roman and Unani. dose of 3, 5 & 7mg/kg body weight orally in the The vast survey of literature showed that form of aqueous slurry. The groups were Ocimum sanctum has a huge spectrum of almost continuously observed for mortality and pharmacological activities. Several research behavioral changes during first 24 hrs and offers evidence that Tulsi is useful against then daily for a fortnight. The observations of stress; it enhances stamina and increases changes in body weight, food and water intake efficient use of oxygen by body; strengthens as well as cage side observations were immune system; reduces inflammation; reported. There was no abnormality observed protects from radiation; reduces aging; in any of these 3 groups. The whole plant supports the lungs, liver and heart; it exhibits powder was found to be nontoxic119. antibiotic, antiviral and antifungal, antioxidant The present study was aimed to study the properties. Different parts of plant have been acute and subacute toxicity studies with orally used in Ayurvedic ancient Medicine to cure an administered 50% ethanolic leaves extract of array of ailments including common cold, Ocimum sanctum Linn (OSE). In acute toxicity cough, headache, flu, asthma, fever, colic tests, four groups of mice (푛 = 6/group/sex) pain, sore throat, bronchitis, hepatic diseases, were orally treated with doses of 200, 600, and malaria fever, flatulence headaches, fatigue, 2000 mg/kg, and general behavior, adverse skin diseases, wound, insomnia, arthritis, effects, and mortality were recorded for up to influenza, digestive disorders, night blindness, 14 days. In subacute toxicity study, rats diarrhea. Tulsi acts as an adaptogen that received OSE by gavage at the doses of 200, helps the body and mind to encounter different 400, and 800 mg/kg/day (푛 = 6/group/sex) for physical, chemical emotional and infectious 28 days, and biochemical, hematological, and stresses, and restore physiological and histopathological changes in tissues (liver, psychological functions. So it can be kidney, spleen, heart, and testis/ovary) were concluded that Ocimum sanctum L. or tulsi is a determined. OSE did not produce any traditionally and clinically proved medicinal hazardous symptoms or death and CNS and herb for both its application and efficacy. ANS toxicities in the acute toxicity test. Subacute treatment with OSE did not show ACKNOWLEDGMENTS any change in body weight, food and water I would like to thank my family and the consumption, and hematological and management of Hindu College of Pharmacy biochemical profiles. In addition, no change for their support and guidance. was observed both in macroscopic and microscopic aspects of vital organs in rats. Our
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